US 12,338,481 B2
Method for producing 2-methyl-butyric acid by bacterial fermentation
Natalia Viktorovna Stoynova, Moscow (RU); Elena Viktorovna Sycheva, Moscow (RU); Anna Alexandrovna Ublinskaya, Moscow (RU); Valery Vasilievich Samsonov, Moscow (RU); and Alexandr Dmitrievich Kivero, Moscow (RU)
Assigned to AJINOMOTO CO., INC., Tokyo (JP)
Filed by AJINOMOTO CO., INC., Tokyo (JP)
Filed on Mar. 24, 2022, as Appl. No. 17/703,519.
Application 17/703,519 is a continuation of application No. PCT/JP2020/035935, filed on Sep. 24, 2020.
Claims priority of application No. RU2019130090 (RU), filed on Sep. 25, 2019.
Prior Publication US 2022/0213515 A1, Jul. 7, 2022
Int. Cl. C12P 7/52 (2006.01); C12N 9/04 (2006.01); C12N 9/10 (2006.01); C12N 9/88 (2006.01)
CPC C12P 7/52 (2013.01) [C12N 9/0006 (2013.01); C12N 9/1025 (2013.01); C12N 9/1096 (2013.01); C12N 9/88 (2013.01); C12Y 101/01085 (2013.01); C12Y 203/03013 (2013.01); C12Y 206/01057 (2013.01); C12Y 402/01033 (2013.01)] 7 Claims
 
1. A method for producing 2-methyl-butyric acid comprising:
(i) cultivating in a culture medium a 2-methyl-butyric acid-producing bacterium belonging to the genus Escherichia to produce and accumulate 2-methyl-butyric acid in the culture medium or cells of the bacterium, or both, and
(ii) collecting 2-methyl-butyric acid from the culture medium or the cells of the bacterium, or both,
wherein said bacterium has been modified to attenuate expression of a gene encoding a protein having tyrosine aminotransferase activity;
wherein said expression of the gene encoding a protein having tyrosine aminotransferase activity is attenuated due to inactivation of the gene,
wherein said gene encoding a protein having tyrosine aminotransferase activity is a tyrB gene, and
wherein said protein having tyrosine aminotransferase activity is selected from the group consisting of:
(A) a protein comprising the amino acid sequence set forth in SEQ ID NO: 2,
(B) a protein comprising the amino acid sequence set forth in SEQ ID NO: 2, comprising substitution, deletion, insertion and/or addition of 1 to 30 amino acid residues, and wherein said protein has tyrosine aminotransferase activity, and
(C) a protein comprising an amino acid sequence having an identity of not less than 90% with respect to the entire amino acid sequence set forth in SEQ ID NO: 2, and wherein said protein has tyrosine aminotransferase activity.